Yeast organisms naturally transport a small number of certain homologous proteins to, and sometimes through, the plasma membrane as an essential contribution to cell surface growth and cell metabolism. As the cell buds as an incident of reproduction preparatory to formation of a daughter cell, additional proteins are required for formation of cell wall and plasma membrane as well as for metabolism. Some of these proteins must find their way to the site of function; hence, a secretory pathway is believed to exist (1). Certain homologous proteins involved in the above processes are formed by translation by ribosomes attached to the endoplasmic reticulum. Homologous proteins are those normally produced by the yeast species and required for its viability. Once formed, they migrate by transfer to Golgi apparatus, thence within vesicles to plasma membranes where some associate, or to some extent, penetrate into the space between the plasma membrane and the cell wall. A small number of homologous proteins seems to be exported completely through the cell wall, such as α-factor and killer toxin (2,3).
Again, the bud region of the cell seems to be the site of attraction for the vesicles and by their fusion to the inner surface of the bud they contribute to the overall growth of the plasma membrane, and presumably, the cell wall (4,5,6). It is controversial still whether glycosylation of the protein may assist, or is implicated, in the so-called secretory process. Further, by definition “secreted” proteins are believed to have a signal prepeptide, postulated to be associated with the transport or incorporation process at the membrane surface. However, the precise mechanism involved in the overall secretory process is not fully understood.
It was contemplated that recombinant DNA technology could provide valuable assistance in answering the open questions about the secretory process in yeast organisms and, given its proven applicability in enabling such, and other, organisms to produce copious quantities of heterologous polypeptide products endogenously (See, e.g., 7 to 17), in achieving appropriate manipulation of the yeast host so as to direct the secretion of heterologous protein in discrete, mature form. This has, in fact, been achieved and is the subject of copending application Ser. No. 06/438,236, and its parent, supra. In that application is described the discovery that a heterologous protein, initially expressed as a pre-protein with its native signal or hybrid thereof, can be processed and secreted by yeast as a mature protein.